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Periodontitis is an infectious disease characterized by the destruction of supporting tissues. Antimicrobial photodynamic therapy (aPDT) has been proposed as an improved method for eliminating microorganisms. Its efficiency depends on the correct use of physical and chemical parameters. Thus, these parameters and their relations were evaluated in this study with the purpose of establishing lethal conditions for combating bacterial agents. Diode lasers and light-emitting diodes (LEDs) were characterized to evaluate the absorption profile and resonance of methylene blue (MB) and toluidine blue O (TBO). The relations between light energy density and photosensitizer absorption were determined. Two methodologies were used to evaluate the effects of aPDT against Aggregatibacter actinomycetemcomitans. LED light exhibited a broad emission spectrum with a peak light wavelength of 637 nm and 99% purity. The resonance intensity of MB was higher with diode laser irradiation, and TBO showed higher resonance intensity with LED irradiation. There was no difference in the absorption profile of photosensitizers using diode lasers or LEDs, and variations in power density did not result in an increasing or decrease in light absorption. A. actinomycetemcomitans was susceptible to photodynamic processes. Emission spectra and peak light wavelengths of light sources combined with the absorption profiles of photosensitizers were the main parameters involved in determining the efficiency of photodynamic effects. Power density did not alter the light absorption of photosensitizers. The association between adequate irradiation characteristics and photosensitizer absorption results in complete inactivation of A. actinomycetemcomitans. In addition, the bactericidal effect was not altered by an increase in energy densities.

Background Bacterial exported proteins represent key components of the host-pathogen interplay. Hence, we sought to implement a combined approach for characterizing the entire exoproteome of the pathogenic bacterium Corynebacterium pseudotuberculosis , the etiological agent of caseous lymphadenitis (CLA) in sheep and goats. Results An optimized protocol of three-phase partitioning (TPP) was used to obtain the C. pseudotuberculosis exoproteins, and a newly introduced method of data-independent MS acquisition (LC-MS E ) was employed for protein identification and label-free quantification. Additionally, the recently developed tool SurfG+ was used for in silico prediction of sub-cellular localization of the identified proteins. In total, 93 different extracellular proteins of C. pseudotuberculosis were identified with high confidence by this strategy; 44 proteins were commonly identified in two different strains, isolated from distinct hosts, then composing a core C. pseudotuberculosis exoproteome. Analysis with the SurfG+ tool showed that more than 75% (70/93) of the identified proteins could be predicted as containing signals for active exportation. Moreover, evidence could be found for probable non-classical export of most of the remaining proteins. Conclusions Comparative analyses of the exoproteomes of two C. pseudotuberculosis strains, in addition to comparison with other experimentally determined corynebacterial exoproteomes, were helpful to gain novel insights into the contribution of the exported proteins in the virulence of this bacterium. The results presented here compose the most comprehensive coverage of the exoproteome of a corynebacterial species so far.

In the present work a family of novel secnidazole-derived Schiff base compounds and their copper(II) complexes were synthesized. The antimicrobial activities of the compounds were evaluated against clinically important anaerobic bacterial strains. The compounds exhibited in vitro antibacterial activity against Bacteroides fragilis, Bacteroides thetaiotaomicron, Bacteroides vulgatus, Bacteroides ovatus, Parabacteroides distasonis and Fusubacterium nucleatum pathogenic anaerobic bacteria. Upon coordination to copper(II) the antibacterial activity significantly increased in several cases. Some derivatives were even more active than the antimicrobial drugs secnidazole and metronidazole. Therefore, the compounds under study are suitable for in vivo evaluation and the microorganisms should be classified as susceptible to them. Electrochemical studies on the reduction of the nitro group revealed that the compounds show comparable reduction potentials, which are in the same range of the bio-reducible drugs secnidazole and benznidazole. The nitro group reduction potential is more favorable for the copper(II) complexes than for the starting ligands. Hence, the antimicrobial activities of the compounds under study might in part be related to intracellular bio-reduction activation. Considering the increasing resistance rates of anaerobic bacteria against a wide range of antimicrobial drugs, the present work constitutes an important contribution to the development of new antibacterial drug candidates.

Coagulase-negative staphylococci (CNS) are important pathogens causing nosocomial infections worldwide with increasing resistance to antimicrobials. The aim of this study was to characterize resistance aspects of CNS isolated from patients with bloodstream infections acquired in hospitals in Belo Horizonte, MG, Brazil. Staphylococcus strains were characterized using repetitive sequence-based polymerase chain reaction (rep-PCR) fingerprinting with (GTG) 5 primer. Phenotypic resistance was analyzed using AST-P5085 card (bioMérieuxVitek ® ). PCR was used to detect mecA , vanA , blaZ , ermA/B/C , aac-aphD , and SCC- mec . For statistical analyses, we used hierarchical cluster, chi-square test (χ 2 ), and correspondence. Several clusters were formed within the same species using (GTG) 5 primer, and strains showed resistance to the following antimicrobials: benzylpenicillin (100%); oxacillin (93.1%); gentamicin (36.3%); ciprofloxacin (63.7%); moxifloxacin (32.7%); norfloxacin (81.0%); erythromycin (86.2%); clindamycin (75.8%); linezolid, teicoplanin and vancomycin (1.7%); tigecycline (0%); fusidic acid (10.35%); rifampicin (13.7%); and trimethoprim/sulfamethoxazole (46.5%). Regarding genotypic analyses, 40%, 0%, 78%, 42%, 100%, 24%, and 30% were positive for mecA , vanA , blaZ , ermA , ermB , ermC , and aac-aphD , respectively. Regarding staphylococcal cassette mec (SCC mec ) type, 3.4% presented type I; 5.0% type II; 27.1% type III; 20.3% type IIIA; and 32.2% type IIIB. Six clusters were formed and frequency distributions of resistant strains to oxacillin, gentamicin, ciprofloxacin, moxifloxacin, norfloxacin, erythromycin, clindamycin, linezolid, teicoplanin, vancomycin, fusidic acid, rifampicin, and trimethoprim/sulfamethoxazole, and mecA , blaZ , ermC , aac-aphD , and SCC mec type differed ( p  < 0.001). In conclusion, the strains investigated in this study were multidrug resistant and carried multiple antibiotic resistance genes.

In this study, phenotypic and genotypic methods were used to detect metallo-β-lactamases, cephalosporinases and oxacillinases and to assess genetic diversity among 64 multiresistant Acinetobacter baumannii strains recovered from blood cultures in five different hospitals in Brazil from December 2008 to June 2009. High rates of resistance to imipenem (93.75%) and polymyxin B (39.06%) were observed using the disk diffusion (DD) method and by determining the minimum inhibitory concentration (MIC). Using the disk approximation method, thirty-nine strains (60.9%) were phenotypically positive for class D enzymes, and 51 strains (79.6%) were positive for cephalosporinase (AmpC). Using the E-test, 60 strains (93.75%) were positive for metallo-β-lactamases (MβLs). All strains were positive for at least one of the 10 studied genes; 59 (92.1%) contained blaVIM-1, 79.6% contained blaAmpC, 93.7% contained blaOXA23 and 84.3% contained blaOXA51. Enterobacteria Repetitive Intergenic Consensus (ERIC)-PCR analysis revealed a predominance of certain clones that differed from each other. However, the same band pattern was observed in samples from the different hospitals studied, demonstrating correlation between the genotypic and phenotypic results. Thus, ERIC-PCR is an appropriate method for rapidly clustering genetically related isolates. These results suggest that defined clonal clusters are circulating within the studied hospitals. These results also show that the prevalence of MDR A. baumannii may vary among clones disseminated in specific hospitals, and they emphasize the importance of adhering to appropriate infection control measures.

Background: Pseudomonas aeruginosa commonly causes nosocomial bloodstream infections and the emergence of a variety of β-lactamases (BLs) is worrying. In 5 hospitals in Belo Horizonte, Brazil, the presence of phenotypes encoding BL genes was established and the genetic diversity of the P. aeruginosa strains recovered from bloodstream infections was analyzed. Materials and Methods: The isolates were investigated using a disk diffusion (DD) method and the Etest®, for encoding metallo-β-lactamases (MBLs), oxacillinases and cephalosporinases. Genes and genetic diversity were evaluated by random amplified polymorphic DNA (RAPD) genotyping and enterobacterial repetitive intergenic consensus (ERIC)-PCR. Results: Twelve strains (30%) were positive for MBLs by Etest and DD, 15 were cephalosporinase-positive and 87.5% were positive for bla SPM-1 and bla VIM-1. Twenty-three strains (57.5%) were grouped into profile A, 32.5% into profile B and 10% into profile C by RAPD genotyping. ERIC-PCR revealed a varying degree of similarity between strains, ranging from 45 to 100%. Conclusions: The results suggest distinct clonal populations in the 5 hospitals studied, indicating a potentially problematic epidemiological situation in Belo Horizonte, Brazil.

Na área rural do município de Teresina-Piauí, foram capturados 129 triatomíneos distribuídos em (a) ecótopos artificiais: habitações (1 Triatoma brasiliensis, 1 Panstrongylus geniculatus, 1 Rhodnius pictipes e 1 Rhodnius prolixusj e galpão abandonado (7 Rhodnius nasutusj e (b) ecótopos naturais: Orbignya martiana (41 Rhodnius neglectus, 33 Rhodnius prolixus e 41 Rhodnius nasutus,) e Copernicia cerifera (3 Rhodnius neglectus,). Cerca de 22,6% dos triatomíneos capturados estavam infectados por flagelados, sendo 30% no ambiente artificial e 21,9% no ambiente natural. Dos 28 mamíferos capturados e examinados, 7 Didelphis aibiventris, 2 Rattus rattus e 1 Tamandua tetradacty la apresentavam-se positivos para flagelados. Os flagelados encontrados nos triatomíneos e nos mamíferos eram semelhantes ao Trypanosoma cruzi. Das 123 reações sorológicas, por imunojluorescência indireta, realizadas na população, duas (1,6%) foram reativas. In the rural areas of Teresina, 129 triatomines were captured distributed in (a) artificial ecotopes; a house with one Triatoma brasiliensis, one Panstrongylus geniqulatus, Rhodniuspictipes, andone Rhodnius prolixus and in a uninhabited chicken house (7 Rhodnius nasutus). (b) Natural ecotopes; Pahus Orbignya martiana (41 Rhodnius neglectus, 33 Rhodnius prolixus and 41 Rhodnius nasutus) and Copernicia cerifera (3 Rhodnius neglectus). The 22.6% of captured triatomines were infected by flagellates similar to Trypanosoma cruzi. Twenty eight sylvatic mammals were captured and examined. Seven Didelphia albiventris, 2 Rattus rattus and a Tamandua tetradactyla were infected withjlagellates. The flagellates found in both triatomines and mammals were morphologically indistinguishable from Trypanosoma cruzi. Serology by the indirect immuno fluorescence testfor Chagas disease revealed two positive seroreactions of positivity among 123 inhabitants examined.

Bacterial exported proteins represent key components of the host-pathogen interplay. Hence, we sought to implement a combined approach for characterizing the entire exoproteome of the pathogenic bacterium Corynebacterium pseudotuberculosis, the etiological agent of caseous lymphadenitis (CLA) in sheep and goats. An optimized protocol of three-phase partitioning (TPP) was used to obtain the C. pseudotuberculosis exoproteins, and a newly introduced method of data-independent MS acquisition (LC-MS.sup.E.sup.) was employed for protein identification and label-free quantification. Additionally, the recently developed tool SurfG+ was used for in silico prediction of sub-cellular localization of the identified proteins. In total, 93 different extracellular proteins of C. pseudotuberculosis were identified with high confidence by this strategy; 44 proteins were commonly identified in two different strains, isolated from distinct hosts, then composing a core C. pseudotuberculosis exoproteome. Analysis with the SurfG+ tool showed that more than 75% (70/93) of the identified proteins could be predicted as containing signals for active exportation. Moreover, evidence could be found for probable non-classical export of most of the remaining proteins. Comparative analyses of the exoproteomes of two C. pseudotuberculosis strains, in addition to comparison with other experimentally determined corynebacterial exoproteomes, were helpful to gain novel insights into the contribution of the exported proteins in the virulence of this bacterium. The results presented here compose the most comprehensive coverage of the exoproteome of a corynebacterial species so far.

Na área rural do município de Teresina-Piauí, foram capturados 129 triatomíneos distribuídos em (a) ecótopos artificiais: habitações (1 Triatoma brasiliensis, 1 Panstrongylus geniculatus, 1 Rhodnius pictipes e 1 Rhodnius prolixusj e galpão abandonado (7 Rhodnius nasutusj e (b) ecótopos naturais: Orbignya martiana (41 Rhodnius neglectus, 33 Rhodnius prolixus e 41 Rhodnius nasutus,) e Copernicia cerifera (3 Rhodnius neglectus,). Cerca de 22,6% dos triatomíneos capturados estavam infectados por flagelados, sendo 30% no ambiente artificial e 21,9% no ambiente natural. Dos 28 mamíferos capturados e examinados, 7 Didelphis aibiventris, 2 Rattus rattus e 1 Tamandua tetradacty la apresentavam-se positivos para flagelados. Os flagelados encontrados nos triatomíneos e nos mamíferos eram semelhantes ao Trypanosoma cruzi. Das 123 reações sorológicas, por imunojluorescência indireta, realizadas na população, duas (1,6%) foram reativas.

Wildfires in humid tropical forests have become more common in recent years, increasing the rates of tree mortality in forests that have not co-evolved with fire. Estimating carbon emissions from these wildfires is complex. Current approaches rely on estimates of committed emissions based on static emission factors through time and space, yet these emissions cannot be assigned to specific years, and thus are not comparable with other temporally-explicit emission sources. Moreover, committed emissions are gross estimates, whereas the long-term consequences of wildfires require an understanding of net emissions that accounts for post-fire uptake of CO2. Here, using a 30 year wildfire chronosequence from across the Brazilian Amazon, we calculate net CO2 emissions from Amazon wildfires by developing statistical models comparing post-fire changes in stem mortality, necromass decomposition and vegetation growth with unburned forest plots sampled at the same time. Over the 30 yr time period, gross emissions from combustion during the fire and subsequent tree mortality and decomposition were equivalent to 126.1 Mg CO2 ha−1 of which 73% (92.4 Mg CO2 ha−1) resulted from mortality and decomposition. These emissions were only partially offset by forest growth, with an estimated CO2 uptake of 45.0 Mg ha−1over the same time period. Our analysis allowed us to assign emissions and growth across years, revealing that net annual emissions peak 4 yr after forest fires. At present, Brazil's National Determined Contribution (NDC) for emissions fails to consider forest fires as a significant source, even though these are likely to make a substantial and long-term impact on the net carbon balance of Amazonia. Considering long-term post-fire necromass decomposition and vegetation regrowth is crucial for improving global carbon budget estimates and national greenhouse gases (GHG) inventories for tropical forest countries.